Ezetimibe is a top‐selling hypolipidemic drug for the treatment of cardiovascular diseases. Biosynthesis of (4S)‐3‐[(5S)‐5‐(4‐fluorophenyl)‐5‐hydroxypentanoyl]‐4‐phenyl‐1,3‐oxazolidin‐2‐one ((S)‐ET‐5) using carbonyl reductase has shown advantages including high catalytic efficiency, excellent stereoselectivity, mild reaction conditions, and environmental friendness, and was considered as the key step for ezetimibe production. The regeneration efficiency of the cofactor, nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) is one of the main restricted factor. Recombinant Escherichia coli strain (smCR125) coexpressing carbonyl reductase (CR125) and glucose dehydrogenase were successfully constructed and applied for the production of (S)‐ET‐5 for the first time. Without extra addition of the coenzyme NADPH, the yield of 99.8% and the enantiomeric excess (e.e.) of 99.9% were achieved under ET‐4 concentration of 200 g/L. Using a substrate fed‐batch strategy, under the optimal conditions, the substrate ET‐4 concentration was increased to 250 g/L with the yield of 98.9% and the e.e. of 99.9% after 12 hr reaction. The space–time yield of 494.5 g L⁻¹ d⁻¹ and the space–time yield per gram biocatalyst of 24.7 g L⁻¹ d⁻¹ g⁻¹ DCW were achieved, which were higher than ever reported for the biosynthesis of the ezetimibe intermediate.

中文翻译：

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使用羰基还原酶与葡萄糖脱氢酶偶联高效生产依折麦布中间体

依折麦布是治疗心血管疾病最畅销的降血脂药物。(4 S )-3-[(5 S )-5-(4-氟苯基)-5-羟基戊酰基]-4-苯基-1,3-恶唑烷-2-酮(( S )-ET-5)的生物合成使用羰基还原酶具有催化效率高、立体选择性好、反应条件温和、环境友好等优点，被认为是依折麦布生产的关键步骤。辅因子烟酰胺腺嘌呤二核苷酸（磷酸）（NAD（P）H）的再生效率是主要的限制因素之一。重组大肠杆菌成功构建共表达羰基还原酶（CR125）和葡萄糖脱氢酶的菌株（smCR125），并首次应用于（S）-ET-5的生产。在不额外添加辅酶 NADPH 的情况下，在 200 g/L 的 ET-4 浓度下实现了 99.8% 的收率和 99.9% 的对映体过量 ( ee )。使用底物分批补料策略，在最佳条件下，反应 12 小时后，底物 ET-4 浓度增加到 250 g/L，产率为 98.9%，ee为 99.9%。494.5 g L ^-1 d ^-1的时空产率和每克生物催化剂24.7 g L ^-1 d ^-1 g ^{-1的时空产率}实现了 DCW，这比以往报告的依折麦布中间体生物合成要高。